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Method for screening lactic acid bacteria in fermented grains of fen-flavor liquor

A technology of fragrance-flavored liquor and screening method, which is applied in the field of liquor brewing to achieve the effects of saving manpower and material resources, being highly targeted, and increasing accuracy

Active Publication Date: 2022-01-11
SHANXI XINGHUACUN FENJIU WINE FACTORY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the technical problem of screening lactic acid bacteria in fermented grains of Fen-flavor liquor, the present invention provides a screening method for lactic acid bacteria with higher selectivity, accuracy and easier operation

Method used

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  • Method for screening lactic acid bacteria in fermented grains of fen-flavor liquor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Screening method for lactic acid bacteria in fen-flavor liquor fermented grains

[0031] 1. Configure the screening medium and the reagents used:

[0032] (1) Amphotericin B and bromocresol green indicator preparation

[0033] Weigh 8 mg of amphotericin B with an analytical balance, put it in a clean bench for ultraviolet sterilization for 15 minutes, then add 1 mL of DMSO to the amphotericin B, mix well, and dispense into 1.5 mL centrifuge tubes to make 8 mg / mL solution.

[0034] Preparation of bromocresol green indicator

[0035] Take 0.1g of bromocresol green and dissolve in 5mL of DMSO, and finally add the prepared solution into 1L of MRS medium.

[0036] (2) Configuration of MRS solid medium

[0037] Peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, glucose 20.0g, Tween-80 1.0mL, diammonium hydrogen citrate 2.0g, anhydrous sodium acetate 5.0g, K 2 HPO 4 2.0g, MnSO 4 ·H 2 O 0.25g, MgSO 4 ·7H 2 O 0.58g, agar 15g, distilled water 1.0L.

[00...

Embodiment 2

[0053] Example 2: Screening of lactic acid bacteria in the fermented grains of Fen-flavor liquor

[0054] (1) The specific implementation method is the same as Example 1, and the difference is that the fermented grains of the Fen-flavor type white wine fermented grains of the vertical fermented grains stage are used to ferment the fermented grains of hawthorn in the late stage;

[0055] The results show:

[0056] 18 strains of lactic acid bacteria were screened from the late stage of fermentation of Fen-flavored liquor fermented grains of Hawthorn in the vertical fermenting stage. After the DNA of the strains was extracted, the DNA chain was replicated by PCR amplification technology. Finally, the strains were identified by 16SrDNA sequencing. Five species of lactic acid bacteria were identified, namely: Lactobacillus plantarum, Lactobacillus coryniformis, Lactobacillus diolivorans, Pediococcus acidilactici, Levy short milk Bacillus (Levilactobacillus brevis).

[0057] The s...

Embodiment 3

[0064] Example 3: Screening of lactic acid bacteria in the fermented grains of Fen-flavor liquor

[0065] The specific implementation method is the same as in Example 1, the difference is that the fermented grains of the Qing-flavor liquor fermented grains of the cold season period and the fermented grains of hawthorn are used in the late stage of fermentation;

[0066] The results show:

[0067] 98 strains of lactic acid bacteria were screened from the late stage of the fermentation of Fen-flavor liquor fermented grains of Dazha in the cold season, and identified by molecular biology (after extracting the DNA of the strains, the PCR amplification technology was used to replicate the DNA chain, and finally the bacteria were identified by 16SrDNA sequencing species) to obtain 9 lactic acid bacteria species, namely: Lactobacillus plantarum (Lactiplantibacillus plantarum), Lactobacillus coryniformis (Lactobacillus coryniformis), Lactobacillus diolivorans, Lactobacillus acidophilu...

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Abstract

The invention discloses a method for screening lactic acid bacteria in fermented grains of fen-flavor liquor, and belongs to the technical field of liquor brewing. The method comprises the following steps: weighing fermented grains, putting the fermented grains into a sterilized PBS buffer solution containing glass beads, and oscillating at room temperature, so that a sample is fully mixed with water; standing after the oscillation is finished, taking supernate and diluting to different gradients; then sucking gradient diluent, uniformly coating the gradient diluent on an MRS culture medium with a bromocresol green indicator for culturing, timely marking out a greening bacterial colony, selecting strains with different forms in the color-changing bacterial colony, and carrying out streak purification on an MRS culture medium plate; and inoculating the single colony into an MRS liquid culture medium, culturing for a certain time, and preserving for later use. The pH value of the culture medium is adjusted to 6.0 by using diluted hydrochloric acid, and amphotericin is added to inhibit fungi. The strain screened by adopting the method has stronger pertinence, the accuracy of lactic acid bacteria screening is increased, and meanwhile, the consumption of manpower and material resources is reduced.

Description

technical field [0001] The invention relates to a method for screening lactic acid bacteria in fermented grains of Fen-flavor liquor, belonging to the technical field of liquor brewing. Background technique [0002] The microbial composition in the fermented grains of Fen-flavor liquor is very complex, mainly including bacteria, molds, and yeasts. The structure of the bacterial flora, especially the lactic acid bacteria flora, plays a decisive role in the quality of the liquor. [0003] Lactic acid bacteria can use fermentable carbohydrates to produce a large amount of lactic acid. Excessive accumulation of lactic acid in the fermented grains will seriously affect the taste of Fen-flavor liquor. has a very important impact. [0004] In the existing related research, the common screening method of lactic acid bacteria is mainly the direct separation of ordinary MRS agar medium, which has low selectivity and is easily interfered by fungi such as yeast and mold; It is time-co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/25C12R1/23C12R1/225C12R1/01
CPCC12N1/20Y02A50/30
Inventor 甄攀赵鑫锐韩英赵欣怡周景文堵国成李江华陈坚蔚慧欣王军燕
Owner SHANXI XINGHUACUN FENJIU WINE FACTORY
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